Magnetic field-loaded lab-on-chip system for the development of cell-free protein synthesis microreactors

• Grant number: 19/00047-3
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: October 01, 2019
• End date: June 30, 2020
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Ricardo Filipe das Neves Lima Pereira
• Grantee: Ricardo Filipe das Neves Lima Pereira
• CNAE: Comércio varejista de outros produtos novos não especificados anteriormente ; Pesquisa e desenvolvimento experimental em ciências físicas e naturais
• Associated researchers: Mario Ricardo Gongora Rubio
• Associated scholarship(s): 20/00447-9 - Lab-on-a-chip system with magnetic field for cell-free protein synthesis microreactors., BP.TT; 19/24048-9 - Lab-on-a-chip system with magnetic field for cell-free protein synthesis microreactors, BP.PIPE

Abstract

In this project, we intend to develop an adequate miniaturized system that incorporates magnetic fields and a lab-on-chip design to be commercialized by BioLinker Synthetic Biology (CNPJ-31.021.329/0001-62). This project came to be out of a need to innovate the process usually used to express and purify recombinant proteins, in order to both shorten it and make it easier as a whole. The cell-free technology (CFT) has been around for awhile and has been put to the aforementioned use in order to increase flexibility in protein synthesis, both in basic and applied research (Carlson et al., 2014). With recent technological advances and a modern approach to modeling new chemical structures application, CFT has had major impact worldwide. Microfluidics is a technology used in advanced manufacturing processes that enables the miniaturization, automation and compartmentalization of the different steps needed to attain a specific goal. In addition, it allows for a significant cost reduction regarding the reagents in a mixture (Li et al., 2014). Last, but not least, there is the possibility of creating a renewable nutrient flow, in order to simulate the behavior of an artificial cell and increasing the yield of production (Dittrich et al., 2005). In this context, we intend to create a lab-on-chip model that optimize the production and synthesis of recombinant proteins which, usually, require 4 distinct and complex steps - biotransformation, clonal selection, expansion and purification of the target molecule. Using microfluidics to integrate transcription and translation, followed by the use of the system already in development in our other project (PIPE fase 1 2017/22801-6) in an integrated system with detection and quantification via biosensors in iterative cycles of protein production is an innovative strategy that will definitely improve on the current standard. Despite current reports on this approach (Georgi et al., 2016; Jackson & Fan, 2007; Marlitt Stech et al., 2014), there are no patents involving such a complete and closed system as the one we are proposing that, in addition to the improvement on the ease of use, also allows for a significant (up to 100x) cost reduction on reagents. In order to do so, the company intends to design the lab-on-chip technology with Prof. Mario Ricardo Gongora Rubio- professor and researcher at the the Manufacture Lab of the Technology Research Institute (IPT). One of the greatest breakthroughs related to the use of microfluidics in microreactors for biotechnological applications is the ease of automation, which leads to high yields in short periods of time, lower production costs with reagents and versatility. (AU)